Analysis of the transcriptional activity of endogenous NFAT5 in primary cells using transgenic NFAT-luciferase reporter mice

Table 2 Hypertonicity threshold required to activate the endogenous 9xNFAT-Luc reporter in transgenic lymphocytes.

		9xNFAT-Luc reporter activity (RLU)
Preactivation with ConA + IL2	Stimulus	Experiment # 1	Experiment # 2	Experiment # 3	Experiment # 4
24 hours	300 mOsm/kg (isotonic)	398	446	761	662
	340 mOsm/kg	295	673	1628	690
	360 mOsm/kg	1254	933	1648	944
	380 mOsm/kg	11468	1622	6060	2353
	400 mOsm/kg	38768	22808	11061	6494
	PMA + ionomycin (isotonic)	7039	20428	12081	6647
48 hours	300 mOsm/kg (isotonic)	257	351	329	210
	340 mOsm/kg	159	397	441	311
	360 mOsm/kg	205	327	567	226
	380 mOsm/kg	1118	1121	1204	1258
	400 mOsm/kg	7105	7997	1891	3291
	PMA + ionomycin (isotonic)	2079	19702	1803	3638
72 hours	300 mOsm/kg (isotonic)	155	356	113	146
	340 mOsm/kg	136	357	131	214
	360 mOsm/kg	128	342	196	99
	380 mOsm/kg	188	379	260	148
	400 mOsm/kg	288	467	458	288
	PMA + ionomycin (isotonic)	11359	22321	2389	1511

Transgenic 9xNFAT-Luc splenocytes were activated with concanavalin A plus IL2 during 24 to 72 hours in isotonic medium, and then exposed to hypertonicity, or stimulated with PMA plus ionomycin for 24 hours. Luciferase activity is represented as relative light units per second (RLU) after normalization with endogenous lactate dehydrogenase.

ISSN: 1471-2199